Applied Microbiology and Biotechnology

, Volume 96, Issue 1, pp 161–170 | Cite as

A new locus affects cell motility, cellulose binding, and degradation by Cytophaga hutchinsonii

  • Xiaofei Ji
  • Yuanxi Xu
  • Cong Zhang
  • Ning Chen
  • Xuemei Lu
Applied genetics and molecular biotechnology
First Online:
Received:
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Accepted:

Abstract

Cytophaga hutchinsonii is a Gram-negative gliding bacterium, which can rapidly degrade crystalline cellulose via a novel strategy without any recognizable processive cellulases. Its mechanism of cellulose binding and degradation is still a mystery. In this study, the mutagenesis of C. hutchinsonii with the mariner-based transposon HimarEm3 and gene complementation with the oriC-based plasmid carrying the antibiotic resistance gene cfxA or tetQ were reported for the first time to provide valuable tools for mutagenesis and genetic manipulation of the bacterium. Mutant A-4 with a transposon mutation in gene CHU_0134, which encodes a putative thiol-disulfide isomerase exhibits defects in cell motility and cellulose degradation. The cellulose binding ability of A-4 was only half of that of the wild-type strain, while the endo-cellulase activity of the cell-free supernatants and on the intact cell surface of A-4 decreased by 40 %. Sodium dodecyl sulfate polyacrylamide gel electrophoresis of proteins binding to cellulose in the outer membrane showed that most of them were significantly decreased or disappeared in A-4 including some Gld proteins and hypothetical proteins, indicating that these proteins might play an important role in cell motility and cellulose binding and degradation by the bacterium.

Keywords

Cytophaga hutchinsonii Gene complementation Cellulose binding protein Cellulose degradation 
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Notes

Acknowledgments

This work was supported by the National Basic Research Program of China (2011CB707402) and the National Natural Science Foundation of China (31170051 and 30870021). We sincerely thank Dr. Mark J. McBride (University of Wisconsin-Milwaukee, Milwaukee, USA) for providing some of the strains and plasmids. Thanks to Dr. Edward C. Mignot, Shandong University, for linguistic advice.

Supplementary material

253_2012_4051_MOESM1_ESM.pdf (43 kb)
ESM 1 (PDF 42 kb)

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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Xiaofei Ji
    • 1
  • Yuanxi Xu
    • 1
  • Cong Zhang
    • 1
  • Ning Chen
    • 1
  • Xuemei Lu
    • 1
  1. 1.State Key Laboratory of Microbial Technology, College of Life ScienceShandong UniversityJinanChina

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